Improved sensitivity of DNA microarrays using photonic crystal enhanced fluorescence.
Improved sensitivity of DNA microarrays using photonic crystal enhanced fluorescence.
复制标题
DOI：
10.1021/ac100841d
复制
发表时间：
2010-08-15
影响因子：
7.4
通讯作者：
Cunningham, Brian T.
中科院分区：
文献类型：
--
作者：
Mathias, Patrick C.;Jones, Sarah I.;Wu, Hsin-Yu;Yang, Fuchyi;Ganesh, Nikhil;Gonzalez, Delkin O.;Bollero, German;Vodkin, Lila O.;Cunningham, Brian T.
DNA微阵列用于以高通量的方式分析样品之间基因表达的变化，但是使用标准微阵列底物测量低表达水平的基因可能会有问题。在这项工作中，我们使用光子晶体（PC）表面扩展了标准微阵列实验的检测能力，该表面增强了从微阵列斑点观察到的荧光。这种PC采用纳米复制成型技术，成本低廉，制造均匀，器件内部和器件之间的光学特性变化很小。通过使用标准协议处理玻璃微阵列底物与pc并行，我们评估了该底物对单色微阵列实验的影响，比较了Glycine max在两个发育阶段的基因表达。PCs将微阵列斑点观察到的信噪比提高了1个数量级，显著增加了底物荧光噪声以上检测到的基因数量。PC底物将分类为差异表达的基因数量增加了一倍以上，甚至可以检测低表达基因的表达变化。这种方法增加了基于表面结合荧光的测定的动态范围，以可靠地定量少量的DNA，这在标准底物中是不可能的。
DNA microarrays are used to profile changes in gene expression between samples in a high-throughput manner, but measurements of genes with low expression levels can be problematic with standard microarray substrates. In this work, we expand the detection capabilities of a standard microarray experiment using a photonic crystal (PC) surface that enhances fluorescence observed from microarray spots. This PC is inexpensively and uniformly fabricated using a nanoreplica molding technique, with very little variation in its optical properties within- and between-devices. By using standard protocols to process glass microarray substrates in parallel with PCs, we evaluated the impact of this substrate on a one-color microarray experiment comparing gene expression in two developmental stages of Glycine max. The PCs enhanced the signal-to-noise ratio observed from microarray spots by 1 order of magnitude, significantly increasing the number of genes detected above substrate fluorescence noise. PC substrates more than double the number of genes classified as differentially expressed, detecting changes in expression even for low expression genes. This approach increases the dynamic range of a surface-bound fluorescence-based assay to reliably quantify small quantities of DNA that would be impossible with standard substrates.
登录
查看更多内容
影响因子：
3.8
作者：
Ganesh, Nikhil;Block, Ian D.;Cunningham, Brian T.
通讯作者：
Cunningham, Brian T.
影响因子：
3.2
作者：
Ganesh, Nikhil;Mathias, Patrick C.;Cunningham, Brian T.
通讯作者：
Cunningham, Brian T.
乙氧基途径的特定元素在幼苗发育过程中大豆子叶的功能过渡中起着重要作用。
影响因子：
4.4
作者：
Gonzalez, Delkin O;Vodkin, Lila O
通讯作者：
Vodkin, Lila O
影响因子：
2.5
作者：
Rosenblatt, D;Sharon, A;Friesem, AA
通讯作者：
Friesem, AA
影响因子：
7.4
作者：
Mathias, Patrick C.;Ganesh, Nikhil;Cunningham, Brian T.
通讯作者：
Cunningham, Brian T.